WO2023025856A1 - Utilisation d'inhibiteurs d'ezh2 pour le traitement de la sténose aortique - Google Patents

Utilisation d'inhibiteurs d'ezh2 pour le traitement de la sténose aortique Download PDF

Info

Publication number
WO2023025856A1
WO2023025856A1 PCT/EP2022/073607 EP2022073607W WO2023025856A1 WO 2023025856 A1 WO2023025856 A1 WO 2023025856A1 EP 2022073607 W EP2022073607 W EP 2022073607W WO 2023025856 A1 WO2023025856 A1 WO 2023025856A1
Authority
WO
WIPO (PCT)
Prior art keywords
ezh2
valve
aortic valve
treatment
aortic
Prior art date
Application number
PCT/EP2022/073607
Other languages
English (en)
Inventor
Sylvain FRAINEAU
Nicolas PERZO
Original Assignee
INSERM (Institut National de la Santé et de la Recherche Médicale)
Centre Hospitalier Universitaire De Rouen
Université De Rouen Normandie
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by INSERM (Institut National de la Santé et de la Recherche Médicale), Centre Hospitalier Universitaire De Rouen, Université De Rouen Normandie filed Critical INSERM (Institut National de la Santé et de la Recherche Médicale)
Priority to EP22765160.1A priority Critical patent/EP4392034A1/fr
Publication of WO2023025856A1 publication Critical patent/WO2023025856A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/507—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/24—Heart valves ; Vascular valves, e.g. venous valves; Heart implants, e.g. passive devices for improving the function of the native valve or the heart muscle; Transmyocardial revascularisation [TMR] devices; Valves implantable in the body
    • A61F2/2412—Heart valves ; Vascular valves, e.g. venous valves; Heart implants, e.g. passive devices for improving the function of the native valve or the heart muscle; Transmyocardial revascularisation [TMR] devices; Valves implantable in the body with soft flexible valve members, e.g. tissue valves shaped like natural valves
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/432—Inhibitors, antagonists
    • A61L2300/434—Inhibitors, antagonists of enzymes

Definitions

  • the present invention is in the field of medicine, in particular cardiology.
  • Aortic valve stenosis also called Calcific aortic valve disease (CAVD)
  • AS also called Calcific aortic valve disease
  • CAVD Calcific aortic valve disease
  • AS also called Calcific aortic valve disease
  • CAVD Calcific aortic valve disease
  • AS also called Calcific aortic valve disease
  • TAVI transcatheter aortic valve replacement
  • VECs Valvular Endothelial Cells
  • Actived monocytes will further undergo differentiation into Ml pro-inflammatory monocytes secreting pro-inflammatory cytokines responsible for inner leaflets Valvular Interstitial Cells (VICs) deactivation (Grim, Aguado et al. 2020) and further differentiation into osteoblast-like cells (Li, Qiao et al. 2017, Raddatz, Huffstater et al. 2020).
  • CAVD progression is due to the imbalance between recruited Mlmonocytes-derived pro-inflammatory macrophages and M2 resident immunomodulatory macrophages populations in the valvular leaflets (Hulin, Hego et al. 2018, Hulin, Hortells et al. 2019, Henaut, Candellier et al. 2019, Karadimou, Plunde et al. 2020).
  • Myeloid cells recruitment (monocytes) and differentiation (into Ml macrophages) represents a critical corner stone during CAVD progression.
  • macrophage phenotype switching from Ml to M2 -like, represents an attractive therapeutic target to design innovative treatments to prevent or cure CAVD patients.
  • Enhancer of zeste homolog 2 (EZH2) part of the Poly comb repressive complex 2 (PRC2) is responsible for generating the H3K27me3 epigenetic modification.
  • TAC Transverse Aortic Constriction
  • cardiac hypertrophy Wang, Zhang et al. 2016, Shi, Fang et al. 2018
  • atrial fibrosis and fibrillation Song, Zhang et al. 2019
  • myocardial infarction Rondeaux et al., under review at Nat Com.
  • the present invention is defined by the claims.
  • the present invention relates to use of EZH2 inhibitors for the treatment of aortic valve stenosis.
  • the inventors show that EZH2 inhibition with GSK-126 and GSK-343 directly regulates monocyte and Ml toward M2 macrophage differentiation, reducing VIC deactivation and osteoblastic transition and thus represents an attractive therapeutic target to prevent CAVD progression.
  • the first object of the present invention relates to a method of treating Aortic valve Stenosis (AS) in a patient in need comprising administering to the patient a therapeutically effective amount of an EZH2 inhibitor.
  • AS Aortic valve Stenosis
  • valve may refer to the valve that prevents the backflow of blood during the rhythmic contractions.
  • aortic valve separates the left ventricle and aorta.
  • the term ’’aortic valve includes a diseased aortic valve or a bioprosthetic valve.
  • bioprosthetic valve is a stented tissue heart valve and may refer to a device used to replace or supplement an aortic valve that is defective, malfunctioning, or missing.
  • bioprosthetic valve prostheses include, but are not limited to, TS 3fs® Aortic Bioprosthesis, Carpentier-Edwards PERIMOUNT Magna Ease Aortic Heart Valve, Carpentier-Edwards PERIMOUNT Magna Aortic Heart Valve, Carpentier-Edwards PERIMOUNT Magna Mitral Heart Valve, Carpentier-Edwards PERIMOUNT Aortic Heart Valve, Carpentier-Edwards PERIMOUNT Plus Mitral Heart Valve, Carpentier-Edwards PERIMOUNT Theon Aortic Heart Valve, Carpentier-Edwards PERIMOUNT Theon Mitral Replacement System, Carpentier-Edwards Aortic Porcine Bioprosthesis, Carpentier-Edwards Dura
  • Aortic Porcine Bioprosthesis Edwards Prima Plus Stentless Bioprosthesis, Edwards Sapien Transcatheter Heart Valve, Medtronic, Freestyle® Aortic Root Bioprosthesis, Hancock® II Stented Bioprosthesis, Hancock II Ultra® Bioprosthesis, Mosaic® Bioprosthesic, Mosaic Ultra® Bioprosthesis, St.
  • bioprosthetic valve comprise a tissue valve having one or more cusps and the valve is mounted on a frame or stent, both of which are typically elastical.
  • the term “elastical” means that the device is able of flexing, collapsing, expanding, or a combination thereof.
  • the cusps of the valve are generally made from tissue of mammals such as, without limitation, pigs (porcine), cows (bovine), horses, sheep, goats, monkeys, and humans.
  • stenosis refers to the narrowing of the aortic valve that could block or obstruct blood flow from the heart and cause a back-up of flow and pressure in the heart.
  • aortic valve stenosis or “AS” or “Calcific aortic valve disease” or “CAVD” has its general meaning in the art and refers to formation, growth or deposition of extracellular matrix hydroxyapatite (calcium phosphate) crystal deposits in the aortic valve.
  • AS aortic valve stenosis
  • CAVD Calcific aortic valve disease
  • treatment refers to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of patient at risk of contracting the disease or suspected to have contracted the disease as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse.
  • the treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment.
  • therapeutic regimen is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy.
  • a therapeutic regimen may include an induction regimen and a maintenance regimen.
  • the phrase “induction regimen” or “induction period” refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease.
  • the general goal of an induction regimen is to provide a high level of drug to a patient during the initial period of a treatment regimen.
  • An induction regimen may employ (in part or in whole) a "loading regimen", which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both.
  • maintenance regimen refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a patient during treatment of an illness, e.g., to keep the patient in remission for long periods of time (months or years).
  • a maintenance regimen may employ continuous therapy (e.g., administering a drug at a regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., disease manifestation, etc.]).
  • the method of the present invention is particularly suitable for primary prevention of AS.
  • the method of the present invention is suitable for the treatment of calcific aortic valve disease.
  • calcific aortic valve disease or “CAVD” has its general meaning in the art and refers to a slowly progressive disorder with a disease continuum that ranges from mild valve thickening without obstruction of blood flow, termed aortic sclerosis, to severe calcification with impaired leaflet motion, or aortic stenosis.
  • disease progression is generally characterized by a process of thickening of the valve leaflets and the formation of calcium nodules — often including the formation of actual bone — and new blood vessels, which are concentrated near the aortic surface.
  • End-stage disease e.g., calcific aortic stenosis
  • End-stage disease is generally characterized pathologically by large nodular calcific masses within the aortic cusps that protrude along the aortic surface into the sinuses of Valsalva, interfering with opening of the cusps.
  • the method of the present invention is particularly suitable for the treatment of calcific aortic stenosis.
  • the method of the present invention is particularly suitable for secondary prevention of aortic valve stenosis.
  • the method of the present invention is particularly suitable for preventing degeneration of an implanted bioprosthetic valve.
  • the method of the present invention is particularly suitable for delaying or preventing the calcification of a bioprosthetic valve after valve replacement either surgically or after Transcatheter Aortic Valve Implantation (TAVI).
  • EZH2 has its general meaning in the art and refers to the enhancer of zeste homolog 2 encoded by the human EZH2 gene (Cardoso, C, et al; European J of Human Genetics, Vol. 8, No. 3 Pages 174-180, 2000).
  • EZH2 is the catalytic subunit of the Polycomb Repressor Complex 2 (PRC2) which functions to silence target genes by tri-methylating lysine 27 of histone H3 (H3K27me3).
  • PRC2 Polycomb Repressor Complex 2
  • H3K27me3 Tri-methylation of H3K27 (H3K27me3) induces chromatin condensation and transcriptional repression of genes involved in development and differentiation (Bracken et al., 2006; Cao et al., 2002; Kirmizis et al., 2004).
  • EZH2 inhibitor refers to a molecule that partially or fully blocks, inhibits, or neutralizes a biological activity or expression of EZH2.
  • the EZH2 inhibitor can be a molecule of any type that interferes with EZH2 in a cell, for example, either by decreasing transcription or translation of EZH2-encoding nucleic acid, or by inhibiting or blocking EZH2 activity, or both.
  • EZH2 inhibitors include, but are not limited to, antisense polynucleotides, interfering RNAs, catalytic RNAs, RNA-DNA chimeras, EZH2-specific aptamers, anti-EZH2 antibodies, EZH2-binding fragments of anti-EZH2 antibodies, EZH2- binding small molecules, EZH2 -binding peptides, and other polypeptides that specifically bind EZH2 (including, but not limited to, EZH2 -binding fragments of one or more EZH2 ligands, optionally fused to one or more additional domains), such that the interaction between the EZH2 inhibitor and EZH2 results in a reduction or cessation of EZH2 activity or expression.
  • EZH2 inhibitors are well known in the art (see e.g. Stazi G, Zwergel C, Mai A, Valente S. EZH2 inhibitors: a patent review (2014-2016). Expert Opin Ther Pat. 2017 Jul;27(7):797-813. doi: 10.1080/13543776.2017.1316976. Epub 2017 Apr 20. PMID: 28394193; Dockerill M, Gregson C, O' Donovan DH. Targeting PRC2 for the treatment of cancer: an updated patent review (2016 - 2020). Expert Opin Ther Pat. 2021 Feb;31(2): 119-135. doi:
  • EZH2 inhibitors include S-Adenosyl-Methionine (SAM)-competitive small molecule inhibitors.
  • SAM S-Adenosyl-Methionine
  • the EZH2 inhibitor is derived from tetramethylpiperidinyl compounds.
  • Further non-limiting examples of EZH2 inhibitors are described in Garapaty-Rao et al., Chemistry and Biology, 20: pp. 1-11 (2013), PCT Patent Application Nos. W02003070887, WO2011140324, WO2011140325, W02012075080, WO2012118812, WO2013/049770, WO2013138361, WO2015104677, WO2015110999, and US Patent Application Nos.
  • the EZH2 inhibitor is Tazemetostat (EPZ-6438) (CAS No. : 1403254- 99-8) having the formula of:
  • the EZH2 inhibitor is GSK-126 ((GSK2816126A) (CAS No. : 1346574- 57-9) having the formula of:
  • the EZH2 inhibitor is GSK-343 (CAS No. : 1346704-33-3) having the formula of:
  • EZH2 inhibitors include ribozymes, antisense oligonucleotides, shRNA molecules and siRNA molecules that specifically inhibit the expression or activity of EZH2.
  • an EZH2 inhibitor comprises an antisense, shRNA, or siRNA nucleic acid sequence homologous to at least a portion of a EZH2 nucleic acid sequence, wherein the homology of the portion relative to the EZH2 sequence is at least about 75 or at least about 80 or at least about 85 or at least about 90 or at least about 95 or at least about 98 percent, where percent homology can be determined by, for example, BLAST or FASTA software.
  • the complementary portion may constitute at least 10 nucleotides or at least 5 nucleotides or at least 20 nucleotides or at least 25 nucleotides or at least 30 nucleotides and the antisense nucleic acid, shRNA or siRNA molecules may be up to 15 or up to 20 or up to 25 or up to 30 or up to 35 or up to 40 or up to 45 or up to 50 or up to 75 or up to 100 nucleotides in length.
  • Antisense, shRNA or siRNA molecules may comprise DNA or atypical or non-naturally occurring residues, for example, but not limited to, phosphorothioate residues.
  • the term "therapeutically effective amount” is meant a sufficient amount of the EZH2 inhibitor for the treatment of the aortic valve stenosis at a reasonable benefit/risk ratio applicable to any medical treatment. It will be understood that the total daily usage of the compound will be decided by the attending physician within the scope of sound medical judgment.
  • the specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific polypeptide employed; and like factors well known in the medical arts.
  • the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day.
  • the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the active ingredient for the symptomatic adjustment of the dosage to the subject to be treated.
  • a medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably from 1 mg to about 100 mg of the active ingredient.
  • An effective amount of the drug is ordinarily supplied at a dosage level from 0.0002 mg/kg to about 20 mg/kg of body weight per day, especially from about 0.001 mg/kg to 7 mg/kg of body weight per day.
  • the EZH2 inhibitor may be combined with pharmaceutically acceptable excipients, and optionally sustained-release matrices, such as biodegradable polymers, to form pharmaceutical compositions.
  • pharmaceutically acceptable excipients such as a carboxylate, a carboxylate, a carboxylate, a carboxylate, a carboxylate, a carboxylate, a carboxylate, a carboxylate, a carboxylate, a carboxysulfate, or adiluent, encapsulating material or formulation auxiliary of any type.
  • the active principle in the pharmaceutical compositions of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal, local or rectal administration, can be administered in a unit administration form, as a mixture with conventional pharmaceutical supports, to animals and human beings.
  • Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual and buccal administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms.
  • the EZH2 inhibitor can be formulated into a composition in a neutral or salt form.
  • Pharmaceutically acceptable salts include the acid addition salts and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like.
  • the carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils.
  • the present invention also relates to the use of an EZH2 inhibitor for the preparation of bioprosthetic valve.
  • the invention relates more particularly to bioprosthetic valve comprising an amount of an EZH2 inhibitor.
  • a local biomaterial or medical delivery device can be used to treat aortic valve stenosis.
  • Such biomaterial or medical delivery device may be biodegradable.
  • the EZH2 inhibitor is preferably entrapped into the cusps of the valve.
  • the cusps of the valve are generally made from tissue of mammals such as, without limitation, pigs (porcine), cows (bovine), horses, sheep, goats, monkeys, and humans. With said entrapment, it is possible to achieve a high level of local action.
  • the valve may be a collapsible elastical valve having one or more cusps and the collapsible elastical valve may be mounted on an elastical stent.
  • the collapsible elastical valve may comprise one or more cusps of biological origin.
  • the one or more cusps are porcine, bovine, or human.
  • the elastical stent portion of the valve prosthesis used in the present invention may be self-expandable or expandable by way of a balloon catheter.
  • the elastical stent may comprise any biocompatible material known to those of ordinary skill in the art.
  • biocompatible materials include, but are not limited to, ceramics; polymers; stainless steel; titanium; nickel -titanium alloy, such as nitinol; tantalum; alloys containing cobalt, such as Elgiloy® and Phynox®; and the like.
  • the process of disposing the coating composition which comprises the EZH2 inhibitor may be any process known in the art.
  • the local delivery according to the present invention allows for high concentration of the EZH2 inhibitor at the disease site with low concentration of circulating compound. For purposes of the invention, a therapeutically effective amount will be administered.
  • FIGURES are a diagrammatic representation of FIGURES.
  • hVICs Human valvular interstitial cells
  • aortic valves form AS patients who underwent aortic valve surgical replacement were collected at Rouen University Hospital, starting on May 2 nd 2016, after agreement of The CPP Nord-rium I (RCB: 2016-A00137-44).
  • Human Valvular Interstitial Cells hVICs were isolated and cultured as already described. (Morvan, Arangalage et al. 2019) Briefly, aortic valve leaflets previously collected and washed in PBS containing antibiotics were cut into pieces and digested with type I collagenase (0.22 U/mg) for 3 hours at 37°C.
  • Digested valves are filters on 70 pm cell strainers to remove undigested valve parts and obtain a homogenous cell suspension resuspended in complete smooth muscle cell basal 2 medium (PromoCell, Heidelberg, Germany) with 10% Fetal Bovine Serum (FBS) Penicillin/Streptomycin (P/S, Sigma-Aldrich Cat#P4333).
  • Cell suspension is then seeded into rat type collagen I (3 mg/ml, Gibco, Cat# A1048301) coated flask and incubated at 37°C, 5% CO2 for several days until reaching cell confluency.
  • hVICs are obtained and cultured in DMEM (Dulbecco’s modified Eagle’s medium, Gibco, Cat# 41966-029) with 10% FBS and P/S until passage 4.
  • DMEM Dulbecco’s modified Eagle’s medium, Gibco, Cat# 41966-029
  • Monocytes were directly isolated from either fresh human CAVD patients (Ref: 2021/229/OB, ID RCB: 2021-A01850-41) or volunteers pooled EDTA peripheral blood samples (Etableau Frangais du Sang, Laboratoire Produits de Laboratoire Enseignement etbericht, Bois capita, France) by negative selection using EasySepTM monocyte isolation kits (StemCell Technologies Cat#19669, Vancouver, Canada) according to the manufacturer’s instructions.
  • Monocytes were then seeded at a cellular density > 150 000 /cm 2 and cultured in DMEM (Gibco, Cat# 41966-029) supplemented with 50 pM of 2-mercaptoethanol (Gibco Cat#21985-023), 10% FBS (Gibco Cat#10500-064), Penicillin/Streptomycin (P/S) and hMacrophage-Colony Stimulating Factor (hM-CSF, 50 ng/ml, StemCell Technologies Cat#78057.1, Vancouver, Canada). Monocytes were differentiated into M0 macrophages after incubation for 4 days with 50 ng/ml hM-CSF.
  • M0 macrophages were further polarized into Ml macrophages after treatment with 50 ng/ml of LipoPolySaccharide (LPS, Sigma-Aldrich Cat#L6529) for 2 days. Selected monocytes and Ml macrophages were then treated for 72 h in the presence of 5 pM EZH2 inhibitors GSK-126 (MedChemExpress, Cat#HY-13470, Monmouth Junction, USA) and GSK-343 (Sigma Aldrich, Cat#SML0766, Saint-Louis, USA) or DiMethyl SulfOxide (DMSO, vehicle).
  • GSK-126 MedChemExpress, Cat#HY-13470, Monmouth Junction, USA
  • GSK-343 Sigma Aldrich, Cat#SML0766, Saint-Louis, USA
  • DMSO DiMethyl SulfOxide
  • CM Conditioned media
  • Previously isolated hVICs were treated for 10 days with human monocytes and Ml macrophages CM diluted v/v with DMEM (Gibco, Cat# 41966-029) in the presence or not of inorganic PyroPhosphate (Pi) at a final concentration up to 1.9 mM (0.9 mM of PPi already present in DMEM). Treatment with conditioned medium was renewed every 2 days and calcium content in each well was measured with the o-cresolphthalein complexone colorimetric method. (Louvet, Buchel et al. 2013, Varennes, Mentaverri et al. 2020) Statistical analyses
  • Data are obtained from indicated number of hVICs isolated from distinct patients. Each o- cresolphthalein complexone measurement experiment was performed in triplicate. Data are expressed as mean values of control (DMSO (vehicle) treated monocytes or Ml macrophages) ⁇ SEM. When indicated, statistical significance was determined by non-parametric Kruskal- Wallis test followed by Dunns post-hoc test using Graphpad Prism 5 software. The use of nonparametric Kruskal-Wallis test was determined depending Shapiro-Wilk normality test data distribution. Data are considered to be significantly different at values p ⁇ 0.05.
  • EZH2 inhibitor GSK-126 is an interesting therapeutic epigenetic treatment to prevent both monocytes and Ml macrophages secretome-induced hVICs calcification and subsequent AS development. We believe that this treatment is of interest to prevent AS development in patients.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Vascular Medicine (AREA)
  • Transplantation (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Dermatology (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La sténose aortique (AS), également appelée rétrécissement aortique calcifié (RAC), est la valvulopathie la plus fréquente en Europe et touche plus de 1 personne de plus de 65 ans sur 4. La progression de l'AS de l'épaississement fibrotique à la calcification des feuillets valvulaires conduit à un développement de l'insuffisance cardiaque et éventuellement à la mort dans les 2 à 5 ans qui suivent l'apparition des symptômes. Les inventeurs montrent maintenant que l'inhibition d'EZH2 avec GSK-126 et GSK-343 régule directement la différenciation des monocytes et des macrophages M1 en macrophages M2, ce qui réduit la désactivation des VIC et la transition ostéoblastique et représente ainsi un objectif thérapeutique intéressant pour empêcher la progression de l'AS. Par conséquent, la présente invention concerne l'utilisation d'inhibiteurs D'EZH2 pour le traitement de la sténose aortique.
PCT/EP2022/073607 2021-08-25 2022-08-24 Utilisation d'inhibiteurs d'ezh2 pour le traitement de la sténose aortique WO2023025856A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP22765160.1A EP4392034A1 (fr) 2021-08-25 2022-08-24 Utilisation d'inhibiteurs d'ezh2 pour le traitement de la sténose aortique

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP21306150 2021-08-25
EP21306150.0 2021-08-25

Publications (1)

Publication Number Publication Date
WO2023025856A1 true WO2023025856A1 (fr) 2023-03-02

Family

ID=77666451

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2022/073607 WO2023025856A1 (fr) 2021-08-25 2022-08-24 Utilisation d'inhibiteurs d'ezh2 pour le traitement de la sténose aortique

Country Status (2)

Country Link
EP (1) EP4392034A1 (fr)
WO (1) WO2023025856A1 (fr)

Citations (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003070887A2 (fr) 2002-02-20 2003-08-28 Sirna Therapeutics, Inc. Inhibition mediee par interference d'arn de l'expression du gene de la proteine ezh2 du groupe polycomb a l'aide d'un acide nucleique court interferent (sina)
US20110251216A1 (en) 2010-02-19 2011-10-13 The Regents Of The University Of Michigan Compositions and methods for inhibiting ezh2
WO2011140325A1 (fr) 2010-05-07 2011-11-10 Glaxosmithkline Llc Indazoles
WO2011140324A1 (fr) 2010-05-07 2011-11-10 Glaxosmithkline Llc Indoles
US20120071418A1 (en) 2010-09-10 2012-03-22 Epizyme, Inc. Inhibitors of Human EZH2 and Methods of Use Thereof
WO2012075080A1 (fr) 2010-12-01 2012-06-07 Glaxosmithkline Llc Indoles
WO2012118812A2 (fr) 2011-02-28 2012-09-07 Epizyme, Inc. Composés hétéroaryles bicycliques substitués condensés en 6,5
WO2013049770A2 (fr) 2011-09-30 2013-04-04 Glaxosmithkline Llc Méthodes de traitement du cancer
WO2013138361A1 (fr) 2012-03-12 2013-09-19 Epizyme, Inc. Inhibiteurs de ezh2 humain et procédés d'utilisation associés
US20140128393A1 (en) 2012-10-15 2014-05-08 Epizyme, Inc. Methods of Treating Cancer
US20140275081A1 (en) 2010-09-10 2014-09-18 Epizyme, Inc. Inhibitors of human ezh2, and methods of use thereof
WO2015104677A1 (fr) 2014-01-10 2015-07-16 Piramal Enterprises Limited Composés hétérocycliques en tant qu'inhibiteurs de ezh2
WO2015110999A1 (fr) 2014-01-24 2015-07-30 Piramal Enterprises Limited Inhibiteurs de ezh2 et leurs utilisations
US20150306281A1 (en) * 2014-04-28 2015-10-29 Nalini Marie Rajamannan Devices and methods for inhibiting stenosis, obstruction, or calcification of a native heart valve, stented heart valve or bioprosthesis
US20160340674A1 (en) * 2014-02-10 2016-11-24 University Of Rochester Compositions and Methods to Inhibit EZH2 for the Treatment of Cardiovascular Diseases
US20210038596A1 (en) * 2018-02-19 2021-02-11 The General Hospital Corporation Methods and compositions for the treatment of vascular disease

Patent Citations (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003070887A2 (fr) 2002-02-20 2003-08-28 Sirna Therapeutics, Inc. Inhibition mediee par interference d'arn de l'expression du gene de la proteine ezh2 du groupe polycomb a l'aide d'un acide nucleique court interferent (sina)
US20110251216A1 (en) 2010-02-19 2011-10-13 The Regents Of The University Of Michigan Compositions and methods for inhibiting ezh2
WO2011140325A1 (fr) 2010-05-07 2011-11-10 Glaxosmithkline Llc Indazoles
WO2011140324A1 (fr) 2010-05-07 2011-11-10 Glaxosmithkline Llc Indoles
US20140275081A1 (en) 2010-09-10 2014-09-18 Epizyme, Inc. Inhibitors of human ezh2, and methods of use thereof
US20120071418A1 (en) 2010-09-10 2012-03-22 Epizyme, Inc. Inhibitors of Human EZH2 and Methods of Use Thereof
WO2012075080A1 (fr) 2010-12-01 2012-06-07 Glaxosmithkline Llc Indoles
WO2012118812A2 (fr) 2011-02-28 2012-09-07 Epizyme, Inc. Composés hétéroaryles bicycliques substitués condensés en 6,5
WO2013049770A2 (fr) 2011-09-30 2013-04-04 Glaxosmithkline Llc Méthodes de traitement du cancer
WO2013138361A1 (fr) 2012-03-12 2013-09-19 Epizyme, Inc. Inhibiteurs de ezh2 humain et procédés d'utilisation associés
US20140128393A1 (en) 2012-10-15 2014-05-08 Epizyme, Inc. Methods of Treating Cancer
WO2015104677A1 (fr) 2014-01-10 2015-07-16 Piramal Enterprises Limited Composés hétérocycliques en tant qu'inhibiteurs de ezh2
WO2015110999A1 (fr) 2014-01-24 2015-07-30 Piramal Enterprises Limited Inhibiteurs de ezh2 et leurs utilisations
US20160340674A1 (en) * 2014-02-10 2016-11-24 University Of Rochester Compositions and Methods to Inhibit EZH2 for the Treatment of Cardiovascular Diseases
US20150306281A1 (en) * 2014-04-28 2015-10-29 Nalini Marie Rajamannan Devices and methods for inhibiting stenosis, obstruction, or calcification of a native heart valve, stented heart valve or bioprosthesis
US20210038596A1 (en) * 2018-02-19 2021-02-11 The General Hospital Corporation Methods and compositions for the treatment of vascular disease

Non-Patent Citations (36)

* Cited by examiner, † Cited by third party
Title
BEGUELIN, W. ET AL.: "EZH2 is required for germinal center formation and somatic EZH2 mutations promote lymphoid transformation", CANCER CELL, vol. 23, no. 5, 2013, pages 677 - 692
BITLER, B. G. ET AL.: "Potential therapeutic targets in ARIDlA-mutated cancers", EXPERT OPIN THER TARGETS, vol. 19, no. 11, 2015, pages 1419 - 1422
BOWDRIDGE, S.W. C. GAUSE: "Regulation of alternative macrophage activation by chromatin remodeling", NAT IMMUNOL, vol. 11, no. 10, 2010, pages 879 - 881
CAMPBELL JEKUNTZ KWKNUTSON SKWARHOLIC NMKEILHACK HWIGLE TJRAIMONDI AKLAUS CRRIOUX NYOKOI A: "EPZ011989, A Potent, Orally-Available EZH2 Inhibitor with Robust in Vivo Activity", ACS MED CHEM LETT, vol. 6, no. 5, 4 March 2015 (2015-03-04), pages 491 - 5, XP055387685, DOI: 10.1021/acsmedchemlett.5b00037
CARDOSO, C ET AL., EUROPEAN J OF HUMAN GENETICS, vol. 8, no. 3, 2000, pages 174 - 180
CAS, no. 1346704-33-3
DAVIS, F. M.K. A. GALLAGHER: "Epigenetic Mechanisms in Monocytes/Macrophages Regulate Inflammation in Cardiometabolic and Vascular Disease", ARTERIOSCLER THROMB VASC BIOL, vol. 39, no. 4, 2019, pages 623 - 634
DOCKERILL MGREGSON CO' DONOVAN DH: "Targeting PRC2 for the treatment of cancer: an updated patent review (2016 - 2020", EXPERT OPIN THER PAT, vol. 31, no. 2, 4 January 2021 (2021-01-04), pages 119 - 135
DUAN RDU WGUO W: "EZH2: a novel target for cancer treatment", J HEMATOL ONCOL, vol. 13, no. 1, 28 July 2020 (2020-07-28), pages 104
GARAPATY-RAO ET AL., CHEMISTRY AND BIOLOGY, vol. 20, 2013, pages 1 - 11
GENTA SPIROSA MCSTATHIS A: "BET and EZH2 Inhibitors: Novel Approaches for Targeting Cancer", CURR ONCOL REP, vol. 21, no. 2, 4 February 2019 (2019-02-04), pages 13, XP036719454, DOI: 10.1007/s11912-019-0762-x
GRIM, J. C. ET AL.: "Secreted Factors From Proinflammatory Macrophages Promote an Osteoblast-Like Phenotype in Valvular Interstitial Cells", ARTERIOSCLER THROMB VASC BIOL, vol. 40, no. 11, 2020, pages e296 - e308
HANAKI SSHIMADA M: "Targeting EZH2 as cancer therapy", J BIOCHEM, 21 January 2021 (2021-01-21), pages mvab007
HENAUT, L. ET AL.: "New Insights into the Roles of Monocytes/Macrophages in Cardiovascular Calcification Associated with Chronic Kidney Disease", TOXINS (BASEL, vol. 11, no. 9, 2019
HSU, A. T. ET AL.: "Epigenetic and transcriptional regulation of IL4-induced CCL17 production in human monocytes and murine macrophages", J BIOL CHEM, vol. 293, no. 29, 2018, pages 11415 - 11423
HULIN, A. ET AL.: "Advances in Pathophysiology of Calcific Aortic Valve Disease Propose Novel Molecular Therapeutic Targets", FRONT CARDIOVASC MED, vol. 5, 2018, pages 21
HULIN, A. ET AL.: "Maturation of heart valve cell populations during postnatal remodeling", DEVELOPMENT, vol. 146, no. 12, 2019
ISHII, M. ET AL.: "Epigenetic regulation of the alternatively activated macrophage phenotype", BLOOD, vol. 114, no. 15, 2009, pages 3244 - 3254, XP002617872
KARADIMOU, G. ET AL.: "TLR7 Expression Is Associated with M2 Macrophage Subset in Calcific Aortic Valve Stenosis", CELLS, vol. 9, no. 7, 2020
LI CWANG YGONG YZHANG THUANG JTAN ZXUE L: "Finding an easy way to harmonize: a review of advances in clinical research and combination strategies of EZH2 inhibitors", CLIN EPIGENETICS, vol. 13, no. 1, 24 March 2021 (2021-03-24), pages 62, XP021288421, DOI: 10.1186/s13148-021-01045-1
LI, G. ET AL.: "The shift of macrophages toward M1 phenotype promotes aortic valvular calcification", J THORAC CARDIOVASC SURG, vol. 153, no. 6, 2017, pages 1318 - 1327, XP085024919, DOI: 10.1016/j.jtcvs.2017.01.052
LIU, Y. ET AL.: "Inhibition of EZH2 attenuates coronary heart disease by interacting with microRNA-22 to regulate the TXNIP/nuclear factor- B pathway", EXP PHYSIOL, vol. 105, no. 12, 2020, pages 2038 - 2050
LOUVET, L. ET AL.: "Magnesium prevents phosphate-induced calcification in human aortic vascular smooth muscle cells", NEPHROL DIAL TRANSPLANT, vol. 28, no. 4, 2013, pages 869 - 878
MORVAN, M. ET AL.: "Relationship of Iron Deposition to Calcium Deposition in Human Aortic Valve Leaflets", J AM COLL CARDIOL, vol. 73, no. 9, 2019, pages 1043 - 1054
NEELE, A. E.M. P. J. DE WINTHER: "Repressing the repressor: Ezh2 mediates macrophage activation", J EXP MED, vol. 215, no. 5, 2018, pages 1269 - 1271
QIAO, Y. ET AL.: "IFN-y Induces Histone 3 Lysine 27 Trimethylation in a Small Subset of Promoters to Stably Silence Gene Expression in Human Macrophages", CELL REP, vol. 16, no. 12, 2016, pages 3121 - 3129
RADDATZ, M. A. ET AL.: "Macrophages Promote Aortic Valve Cell Calcification and Alter STAT3 Splicing", ARTERIOSCLER THROMB VASC BIOL, vol. 40, no. 6, 2020, pages el53 - el65
SATOH, T. ET AL.: "The Jmjd3-Irf4 axis regulates M2 macrophage polarization and host responses against helminth infection", NAT IMMUNOL, vol. 11, no. 10, 2010, pages 936 - 944, XP055025818, DOI: 10.1038/ni.1920
SHI, Z. L. ET AL.: "EZH2 Inhibition Ameliorates Transverse Aortic Constriction-Induced Pulmonary Arterial Hypertension in Mice", CAN RESPIR J, vol. 2018, 2018, pages 9174926
SONG, S. ET AL.: "EZH2 as a novel therapeutic target for atrial fibrosis and atrial fibrillation", J MOL CELL CARDIOL, vol. 135, 2019, pages 119 - 133, XP085815691, DOI: 10.1016/j.yjmcc.2019.08.003
STAZI GZWERGEL CMAI AVALENTE S: "EZH2 inhibitors: a patent review (2014-2016", EXPERT OPIN THER PAT., vol. 27, no. 7, 20 April 2017 (2017-04-20), pages 797 - 813
VAN DEN BOSSCHE, J. ET AL.: "Macrophage polarization: the epigenetic point of view", CURR OPIN LIPIDOL, vol. 25, no. 5, 2014, pages 367 - 373
VARENNES, O. ET AL.: "The Metabolism of Epoxyeicosatrienoic Acids by Soluble Epoxide Hydrolase Is Protective against the Development of Vascular Calcification", INT J MOL SCI, vol. 21, no. 12, 2020
WANG, Z. ET AL.: "The long noncoding RNA Chaer defines an epigenetic checkpoint in cardiac hypertrophy", NAT MED, vol. 22, no. 10, 2016, pages 1131 - 1139
WEI, X. ET AL.: "Pharmacological inhibition of EZH2 by GSK126 decreases atherosclerosis by modulating foam cell formation and monocyte adhesion in apolipoprotein E-deficient mice", EXP THER MED, vol. 22, no. 2, 2021, pages 841
ZHANG, X. ET AL.: "Macrophage/microglial Ezh2 facilitates autoimmune inflammation through inhibition of Socs3", J EXP MED, vol. 215, no. 5, 2018, pages 1365 - 1382, XP055945054, DOI: 10.1084/jem.20171417

Also Published As

Publication number Publication date
EP4392034A1 (fr) 2024-07-03

Similar Documents

Publication Publication Date Title
AU2018254743B2 (en) Combination comprising a PPAR agonist such as elafibranor and an acetyl-CoA carboxylase (ACC) inhibitor
JP2020521734A (ja) 老化細胞除去化合物
Giordano et al. Convertible visceral fat as a therapeutic target to curb obesity
RU2426532C2 (ru) Применение органических соединений
Lee et al. Sulfuretin, a major flavonoid isolated from Rhus verniciflua, ameliorates experimental arthritis in mice
JP7548567B2 (ja) Ｉｌ－３３を含有するマトリックス結合型小胞（ｍｂｖｓ）およびそれらの使用
ES2869463T3 (es) Composición para la prevención o tratamiento de la calcificación de válvulas que contiene un inhibidor de DPP-4
Jeon et al. Inhibitory effects of PPARγ ligands on TGF-β1-induced CTGF expression in cat corneal fibroblasts
Lee et al. Ezetimibe ameliorates lipid accumulation during adipogenesis by regulating the AMPK–mTORC1 pathway
Yang et al. Myeloid-derived growth factor suppresses VSMC dedifferentiation and attenuates postinjury neointimal formation in rats by activating S1PR2 and its downstream signaling
US20240350508A1 (en) Use of ezh2 inhibitors for the treatment of aortic valve stenosis
MX2007015083A (es) Combinacion de inhibidores de hmg-coa-reductasa e inhibidores de mtor.
WO2023025856A1 (fr) Utilisation d'inhibiteurs d'ezh2 pour le traitement de la sténose aortique
Biller et al. Effect of metformin on HIF-1α signaling and postoperative adhesion formation
TWI344366B (en) Pharmaceutical compositions for the treatment of renal dysfunction, disease or disorder, in particular in diabetic patients
US20110015410A1 (en) Novel use of scoparone
KR102350697B1 (ko) 페오포르바이드계 화합물을 유효성분으로 포함하는 섬유증의 예방 또는 개선용 식품 조성물
Li et al. CREG: a possible candidate for both prevention and treatment of proliferative vascular disease
AU2004231345B2 (en) Use of steroid derivatives for the treatment of angiotensin II related disease e.g. cardiovascular and proliferative disorders
WO2012105706A1 (fr) Agent thérapeutique contre les chéloïdes
KR20160062517A (ko) Ｄｐｐ­４ 저해제를 유효성분으로 함유하는 혈관 석회화 억제용 약학적 조성물
Mondin et al. Alterations in cytoskeletal protein expression by mycophenolic acid in human mesangial cells requires Rac inactivation
Brietzke et al. Actinomycin D for fibrosis management in ophthalmic implant surgery
Meng et al. Chinese rice wine polyphenol compounds inhibit vascular smooth muscle cell dedifferentiation and its mechanism
Zang et al. Protective Effect of CD137 Deficiency against Post-infarction Cardiac Fibrosis and Adverse Cardiac Remodeling via ERK1/2 Signaling Pathways

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 22765160

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2022765160

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2022765160

Country of ref document: EP

Effective date: 20240325